LOCUS       sequence-77281-e         927 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 927)
  AUTHORS   Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E, 
            Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA
  TITLE     Design of orthogonal genetic switches based on a crosstalk map of 
            sigmas, anti-sigmas, and promoters.
  JOURNAL   Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58.
  PUBMED    24169405
REFERENCE   2  (bases 1 to 927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2013-10-29"; volume: "9:702. doi"; pages: " 
            10.1038/msb.2013.58"
COMMENT     Sequence Label: pVRa34_3302
FEATURES             Location/Qualifiers
     source          1..927
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    80..104
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             119..141
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             160..177
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             187..210
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             808..825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(878..896)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 gcggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
       61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
      121 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      181 agcggcctgg aagttctgtt ccaggggccc catatggatg ataccgatct gattgatcgt
      241 gttgcacgtg ccgatccgaa agcactgggt catctgtatg attgttatgc accgctgctg
      301 ctggcagttg gtctgggtat tctgagcaat cgtcaagagg ccgaagatgt tctgcataat
      361 gtgtttatgg aaatttggcg taaagccagc cagtatgatg gtcgtcgtgg tagcgttcgt
      421 gcatggctga ttctgcgtat gcgtagccgt agcctggatc gtctgcgtag cgcaccgtat
      481 acccgtagca ttccgattcc ggaaaccgtt ctggaaaata ccgcaaatgg tatgagccaa
      541 gatccggcac tggcatggga ttgtcaggca gttcgtaaag caatgagcaa actgagccgt
      601 gaacagcgta ccattatgga actggcatat tttcagggtc tgagcgcaag cgaaattgca
      661 agccatgttg gtctgccgct gggcaccatt aaaagccgta tgcgtggtgc actgaatgtt
      721 ctgcgtaccc attatcgtac ccagtgtcag cgtattagca atgaaaccga aggcaaataa
      781 ggatccaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct
      841 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      901 ccccttgggg cctctaaacg ggtcttg
//
